Pharmaceutical and diagnostic use of Serum Amyloid P component

ABSTRACT

The invention relates to the new use of Serum Amyloid P component (SAP) and/or endotoxin-binding fragments thereof for the preparation of a pharmaceutical composition for neutralizing lipopolysaccharide and particularly for the treatment of sepsis. The invention further relates to a diagnostic method for demonstrating the presence of endotoxin in blood or blood fractions, such as serum or plasma.

This application is a 371 of PCT/NL97/00567 filed on Oct. 10, 1997.

The present invention relates to a new method of treating sepsis. Theinvention further provides possibilities for therapeutic and preventivetreatment of Alzheimer's disease.

Sepsis very generally comprises the clinical pictures which result fromthe presence of bacteria multiplying in blood. The direct cause of thesymptoms are toxic substances which are released by the bacteria orreleased during lysis thereof. Gram-negative bacteria for instanceproduce lipopolysaccharides (LPS) as a component of their cell wall.These lipopolysaccharides are toxic in many circumstances. In principlethey are bound to the cell and are only released when the cell lyses.Lipopolysaccharides are also referred to as endotoxins.

Infection with Gram-negative bacteria can result in a life-threateningdisease which is initiated by specific binding of LPS to phagocytes,such as monocytes, macrophages and granulocytes (neutrophils). These arehereby activated and secrete various cytokines, such as tumor necrosefactor-α (TNF-α), interleukin 1 (IL-1), IL-6, IL-8, and otherinflammation mediators. These compounds initiate a cascade of events,either directly or by activation of secondary mediators, whichultimately result in fever and disorders in the coagulation of theblood, vasodilation, organ failure and finally septic shock.

Different treatments for sepsis have already been proposed. Thus, forthe treatment of specific Gram-negative sepsis, monoclonal antibodiesare for instance used against endotoxin. Tests are currently also beingcarried out with recombinant BPI, a product of the neutrophils with astrong lipopolysaccharide-neutralizing effect.

Used for treatment of general sepsis are antibodies aimed againstcytokines or antagonists for the soluble TNF (tumor necrose factor)receptor or for the interleukin-1 receptor.

Up to the present a totally satisfactory result has not yet beenachieved with the known methods.

It is therefore the first object of the present invention to provide anew method of treatment and diagnosis of sepsis.

Surprisingly, it has been found that the per se known protein SerumAmyloid P component (SAP) is capable of binding to endotoxin. Binding ofthe circulating SAP to the phagocytes, and therewith activation thereof,is hereby prevented. In this manner SAP is capable of neutralizing thebiological action of endotoxin.

The present invention therefore relates in a first aspect to the use ofSerum Amyloid P component (SAP) for the preparation of a pharmaceuticalcomposition for neutralizing lipopolysaccharide(s) in general and thetreatment of sepsis in particular.

SAP is a member of the family of the pentraxins. Pentraxins are proteinswith a characteristic pentameric organization of identical subunitswhich are ordered as single or double annular discs. Another member ofthis family is the C-reactive protein (CRP). CRP and SAP are bothso-called “acute-phase reactants” (APR), i.e. they are involved in theearly phase of an inflammation process. Per species however, it isgenerally found that only one of the two acts as APR. For humans it isthe case that the concentration of SAP in normal human plasma isapproximately 30 μg/ml. During inflammation reactions this level remainsroughly the same, while the CRP level may well be increased athousand-fold to 1 mg/ml, depending on the disease and the seriousnessthereof.

It may be considered particularly surprising that a compound which playsno part as APR during inflammation reactions is in fact capable ofneutralizing sepsis-causing endotoxin. It could be expected that thebody itself would make use of this neutralizing capacity by increasingthe plasma level of this compound in the case of infection withGram-negative bacteria and the inflammation reactions resultingtherefrom.

SAP is a relatively large protein. For particular ligands is known whichof the amino acids of SAP are involved in binding, such as for instancefor CRP, C4-binding protein (C4bp), Clq and Calcium ions. It has thusalso been established for endotoxin that determined regions of the SAPmolecule are involved in the binding. It may therefore be recommended touse only the specific binding parts of the protein for manufacture of apharmaceutical preparation.

According to the invention it has now further been found that SAPfragments, which consist of at least a part of the amino acids 27-39 ofSAP in the sequence occurring in SAP, are very successful in inhibitingthe binding of LPS to monocytes.

According to a second aspect of the invention new peptides (SAPfragments) are therefore provided for use in neutralizinglipopolysaccharides, which peptides consist of a part of the amino acids27-39 of SAP in the sequence occurring in SAP. Preferred peptides arethe PEP 27-39, which consists of the amino acids 27-39, PEP 33-38 (aminoacids 33-38), PEP 32-39 (amino acids 32-39), PEP 30-37 (amino acids30-37) and PEP 29-36 (amino acids 29-36). Table 1 below shows the aminoacid composition of the different peptides.

TABLE 1 Amino Acid Number 27 28 29 30 31 32 33 34 35 36 37 38 39 SEQ IDNO SAP PEP 29-39 Glu Lys Pro Leu Gln Asn Phe Thr Leu Cys Phe Arg Ala 1PEP 33-38 Phe Thr Leu Cys Phe Arg 2 PEP 32-39 Asn Phe Thr Leu Cys PheArg Ala 3 PEP 33-38 Gln Asn Phe Thr Leu Cys Phe Arg 6 PEP 30-37 Leu GlnAsn Phe Thr Leu Cys Phe 4 PEP 29-36 Pro Leu Gln Asn Phe Thr Leu Cys 5PEP 28-35 Lys Pro Leu Gln Asn Phe Thr Leu 7 PEP 27-34 Glu Lys Pro LeuGln Asn Phe Thr 8 Glu-Glutamate; Lys-Lysine; Pro-Proline; Leu-Leucine;Gln-Glutamine; Asn-Asparagine; Phe-Phenylalanine; Thr-Threonine;Cys-Cysteine; Arg-Arginine; Ala-Alanine

The advantage of SAP fragments is that they can be manufactured moresimply owing to their smaller dimensions and can penetrate more easilyinto bodily tissues.

In the research which led to the present invention it was furtherinferred that LPS is possibly involved as environmental factor in thedevelopment of Alzheimer's disease. Alzheimer's disease coincides withparticular forms of cancer, rheumatoid arthritis, diabetes and Down'ssyndrome under the denominator ‘amyloidosis’. This is a collection ofdiseases which are characterized by extracellular deposits of normal ormutated proteins. The amyloid deposits in Alzheimer's are ordered in acharacteristic three-dimensional pattern of so-called “beta-pleatedsheets”. The subunit protein component consists of the amyloidbeta-protein (A beta-P). This is a small fragment of approximately 40amino acids which is released by enzymes from the transmembranebeta-amyloid precursor protein (beta-APP). The processing of thisprecursor protein can take place in a number of ways and then results ina normally occurring soluble fragment or, under certain conditions viaalternative proteases, an intact beta-fragment. The production ofamyloid beta-protein is therefore in itself a normal physiological eventand the existence of A beta-P can be demonstrated in the cerebral fluid(CFS) of healthy humans. However, the deposit of amyloid beta-protein isthe primary event causing Alzheimer's disease.

Other proteins are also associated with the amyloid deposits, includingSAP and serum amyloid A (SAA). According to the invention it has nowbeen found that both SAP and SAA can bind to LPS and are capable ofneutralizing the biological activity of LPS. As such these two amyloidassociated plasma proteins have no structural affinity.

According to the present invention it is now proposed that LPS entersinto a binding with the serum amyloid proteins SAP and SAA, whereby therole of SAP and SAA in the initiation of amyloid deposits is influenced.It is suspected that through binding of LPS to SAP and SAA theoccurrence of deposits is stimulated. The hypothesis now is that(chronic) bacterial infections, and particularly LPS as environmentalfactor, contribute to the development of Alzheimer's disease. It is infact suspected that the basis for Alzheimer's is the alternativeprocessing of the beta-amyloid precursor protein, which possibly takesplace outside the brain in the circulation. There are indications thatsoluble amyloid beta-protein in plasma is associated with lipoproteins,in particular the VHDL and HDL3 fractions, in which it is complexed withthe apolipo protein J (ApoJ). The ApoJ/amyloid beta-protein complex iscapable of passing through the blood-brain barrier. In this manner theamyloid beta-protein enters the brain, where it is deposited. SAA islikewise an apolipo protein which is associated with the HDL fractionand particularly with the HDL3 subfraction.

It is further suspected that LPS also plays a part in the development ofAlzheimer's disease via an indirect route. Cytokines, such asinterleukin 1 (IL-1) and interleukin 6 (IL-6), lead to over-expressionof beta-amyloid precursor protein in the vessel wall and in microgliaand astrocytes in the brain. This points to a role for the acute-phaseresponse in the development of Alzheimer's disease. LPS is a potentinitiator of the acute-phase response and also initiates the productionof IL-1 and IL-6. Because both cytokines result in more beta-amyloidprecursor protein, an indirect role of LPS is assumed.

The SAP itself and fragments derived from SAP (peptides) with a strongLPS-binding and neutralizing action can therefore be of importance ineliminating the part played by LPS in the development of Alzheimer'sdisease.

This influence can relate to the initiation as well as the furtherprogression of the disease.

According to a third aspect the present invention therefore provides theuse of SAP and/or SAP fragments thereof for the manufacture of apharmaceutical composition for the therapeutic and preventive treatmentof Alzheimer's disease.

The pharmaceutical compositions, which according to the inventioncontain SAP and/or one or more SAP fragments (peptides) as activeingredient, will be particularly intended for parenteral, and thenparticularly intravenous use. The pharmaceutical compositions can beprepared by combining (i.e. mixing dissolving etc.) SAP and/or one ormore SAP fragments with pharmaceutically acceptable excipients suitablefor intravenous administration. The concentration of the activeingredient in a pharmaceutical composition can vary between 0.001% and100%, depending on the nature of the treatment and the manner ofadministration. The dose of the active ingredient to be administeredlikewise depends on the administering route and application but can forinstance vary between 0.01 μg and 1 mg per kg body weight, preferablybetween 0.1 μg and 100 μg per kg body weight.

In addition to use in a pharmaceutical composition, SAP and/or the SAPfragments can also be used for diagnosis of infection with Gram-negativebacteria or sepsis. For this purpose the invention provides a diagnosticmethod for demonstrating the presence of endotoxin in blood or bloodfractions, such as serum or plasma, comprising of bringing a carrierwith SAP and/or endotoxin-binding SAP fragments bound thereto intocontact with a blood sample for testing in order to enable binding ofendotoxin to SAP(-fragments), removing non-bound material andvisualizing and/or quantifying the binding between endotoxin andSAP(-fragments).

The invention further provides a diagnostic kit for performing themethod, comprising a carrier having bound thereto SAP and/orendotoxin-binding fragments thereof, and means for visualizing and/orquantifying binding between endotoxin and SAP(-fragments).

The carrier can take different forms, such as a microtitre plate, acolumn, a membrane or beads. These latter can for instance be magneticbeads, such as Dyna-beads™.

Binding of endotoxin to SAP or SAP fragments can be detected indifferent ways. Use can thus be made of a labelled antibody againstendotoxin.

In the present application the terms ‘SAP fragment(s)’, ‘peptide(s)’ and‘endotoxin-binding peptide(s)’ are used interchangeably.

The binding of SAP to endotoxin was found by “fishing” in plasma withmagnetic beads having endotoxin bound thereto. The neutralizing actionwas tested on monocytes, neutrophils and in mice. In addition, SAPfragments were manufactured and the neutralizing action thereof wastested on monocytes. Details of these tests are described in theexamples following hereinbelow, which are only given by way ofillustration and are not intended to limit the invention in any waywhatever.

Reference will be made in the examples to the following accompanyingfigures:

FIG. 1A shows an SDS-PAGE gel of LPS-Beads which are incubated withserum pre-incubated with free LPS.

lane 1: markers (respectively from top to bottom 92, 66, 45, 31 an d21kDa),

lane 2: without pre-incubation with free LPS,

lane 3: pre-incubation with 100 μg/ml free LPS

lane 4: pre-incubation with 10 μg/ml free LPS

lane 5: pre-incubation with 1 μg/ml free LPS.

FIG. 1B shows an SDS-PAGE gel of LPS-Beads incubation with serum (lane1, negative control), commercially obtained SAP (lane 2) and markers(lane 3, molecular weight respectively from top to bottom 92, 66, 45, 31and 21 kDa).

FIG. 2 shows the effect of SAP on LPS-binding to monocytes. Plotted onthe X-axis is the concentration of SAP. The Y-axis shows the averagefluorescence which represents the ReLPS-binding.

FIG. 3 shows the inhibition of the LPS priming of neutrophils by SAP inthe presence and absence of LPS. In the graph the luminescence countsare plotted against the measurement time (in minutes).

FIG. 4 shows the effect of peptides (SAP fragments) according to theinvention on the binding of LPS to monocytes. Plotted on the X-axis isthe concentration of the peptides. The Y-axis shows the averagefluorescence, which represents the ReLPS-binding.

EXAMPLES Example 1 Identification and isolation of SAP asendotoxin-binding molecule

1.1 Materials and method A 1 mg/ml solution of LPS of Salmonella Re595(Sigma; L9764) in carbonate buffer (0.1 M, pH 9.5) is incubated for 24hours with 5×10⁷ DynaBeads™ (M450, Dynal A. S., Oslo, Norway) activatedwith Tosyl. The beads loaded with LPS are washed 3x and stored inHBSS+0.1% Na-azide (5×10⁷ b/ml). A 10% dilution (1 to 10 dilution) ofserum of healthy volunteers is mixed with 5×10⁶ LPS-Beads for 30 minutesat 22° C. The serum has optionally been incubated before hand withdifferent concentrations of free LPS. The beads are washed 3x with HBSS(Gibco BRL, Gaithersburg, Md., US) and subsequently resuspended in 25 μlSDS-PAGE sample buffer (with 2% SDS and 2.5% DTT). After boiling for 3minutes at 100° C. the sample is analyzed on a 12.5% SDS-PAGE(Mini-ProteanII; Bio-Rad). The proteins present are stained withCoomassie Blue. A sample with Marker proteins of known sizes (Bio-Rad)serves as reference. For identification the proteins are transferredafter SDS-PAGE to nitrocellulose paper by means of a MiniTrans-Blotter™. The proteins are stained with Coomassie Blue, and theprotein around 30 kDa is excited and dissolved in buffer. The sequenceof this sample is determined. SAP is isolated from normal human serum bymaking use of the Ca-dependent binding to Sepharose. With delipidatedserum, which is manufactured by centrifuging serum for 4 minutes at16,000 rpm, two precipitation steps are first performed (successivelywith BaCl₂ and NH₄SO₄) followed by anion exchange chromatography on aMono-Q column (Pharmacia). The eluate at 0.2 to 0.3 M NaCl is finallyguided over a Sepharose-4B column and 95% pure SAP is obtained byelution with EDTA (analysis on SDS-PAGE and with a specific ELISA). Forthe purification method see also Skinner & Cohen, Methods in Enzymology,vol. 163, pages 523-536 (1988).

1.2 Results

1. FIG. 1A shows an example of a SDS-PAGE gel of LPS-beads incubatedwith serum. For specificity of the reaction the serum was firstincubated with different concentrations (100, 10, 1 and 0 μg/ml) of freeReLPS (L964, Sigma Chemicals, St. Louis, Mo. US) before the mixture wasincubated with the LPS-Beads. This demonstrates that when SAP issaturated with free LPS no further binding to the LPS-Beads occurs(lanes 3 and 4). It follows herefrom that SAP is also capable of bindingwith free LPS.

2. Sequence determination of the first 17 amino acids (N-terminal) ofthe ±30 kDa protein resulted in a sequence which corresponds 100% withSerum Amyloid P-Component (SAP). The probability that the protein is SAPis hereby 89%. This protein and SAP were used to demonstrate theneutralizing action thereof.

3. Binding of commercially obtained SAP (Calbiochem; 565190) toLPS-Beads by analysis with SDS-PAGE is shown in FIG. 1B. This again alsoshows that the component from serum which binds to the LPS-Beads isindeed SAP.

Example 2 Inhibition of endotoxin-induced monocyte activation by SAP

2.1 Materials and method LPS of Salmonella Re595 (ReLPS, supra) islabelled with FITC (Sigma; F7250) under conditions in which LPS ispresent as monomer, resulting in a FITC-LPS preparation with a ratio of1 molecule FITC per molecule PLS. After extension desalting and dialysisagainst PBS, the stock solution of FITC-LPS is stored at −20° C.Mononuclear leukocytes (monocytes and lymphocytes) are isolated fromheparin blood of healthy volunteers via a Ficol (Pharmacial) gradient inaccordance with a standard method. After washing the cells areresuspended in isotonic HEPES buffer (with 1 mM CalCl₂ and 2% BSA).Preparations with SAP are mixed with FITC-LPS and subsequently added to3×10⁵ mononuclear cells and 30 ng/ml recombinant LSP (obtained from Dr.Lichtenstein) in a total volume of 50 μl and a FITC-LPS concentration of2.5 ng/ml. After an incubating period of 30 minutes at 37° C., thesamples are stored on ice. Analysis of monocyte-associated FITC-LPSbinding takes place with PAC Scan™ Flow cytometer. Forward and SidewardScatter parameters are used to identify the monocyte population. WithLysisII™ software (Beckton & Dickinson) the average fluorescence valueof 5000 monocytes is calculated. See also Weersing A. J. L. et al., J.Immunol. 145, 318-324 (1990)).

2.2 Results

1. FIG. 2 shows the concentration-dependent inhibition of FITC-LPSbinding to monocytes in vitro. This shows that the more SAP is added inthe assay, the less LPS associates itself with the monocytes. Sinceassociation of LPS with the monocyte is the first step in the activationof the monocyte, this is a very strong indication that SAP will alsoinhibit the monocyte activation in vivo at a very early stage.

Example 3 Inhibition of endotoxin-induced monocyte activation by SAPfragments

3.1 Materials and method The test was performed in the same manner asdescribed in example 2. However, SAP fragments (peptides) were usedinstead of SAP. The amino acid composition of the tested peptides isshown in table 2, as well as the concentration at which 50% inhibitionoccurs. FIG. 4 shows the inhibition at different concentrations. ‘PEP27-39scr’ herein represents a peptide with the same amino acidcomposition as PEP 27-39, but in a different sequence.

3.2 Results Table 2 and FIG. 4 show that particularly PEP 27-39 has avery good activity. PEP 33-38, PEP 32-39, PEP 30-37 and PEP 29-36 alsostill have an acceptable activity. The result with PEP 27-39 scr showsthat not only the amino acid composition but also the amino acidsequence have an influence on the inhibiting action of a peptide onbinding of LPS to monocytes.

TABLE 2 Amino Acid 50% inhibition SEQ ID Number 27 28 29 30 31 32 33 3435 36 37 38 39 concentration NO SAP 0.05 μM PEP 29-39 Glu Lys Pro LeuGln Asn Phe Thr Leu Cys Phe Arg Ala 0.1 μM 1 PEP 33-38 Phe Thr Leu CysPhe Arg 3.5 μM 2 PEP 32-39 Asn Phe Thr Leu Cys Phe Arg Ala 2 μM 3 PEP33-38 Gln Asn Phe Thr Leu Cys Phe Arg >100 μM 6 PEP 30-37 Leu Gln AsnPhe Thr Leu Cys Phe 4.5 μM 4 PEP 29-36 Pro Leu Gln Asn Phe Thr Leu Cys6.5 μM 5 PEP 28-35 Lys Pro Leu Gln Asn Phe Thr Leu >100 μM 7 PEP 27-34Glu Lys Pro Leu Gln Asn Phe Thr >106 μM 8 Glu-Glutamate; Lys-Lysine;Pro-Proline; Leu-Leucine; Gln-Glutamine; Asn-Asparagine;Phe-Phenylalanine; Thr-Threonine; Cys-Cysteine; Arg-Arginine;Ala-Alanine

Example 4 Inhibition of endotoxin-induced neutrophil activation by SAP

4.1 Materials and method Neutrophils are isolated from heparin blood ofhealthy volunteers via a Histopaque-Picoll gradient in accordance with astandard method (Van Amerafoort, E. S. & J. A. G. van Strijp, Cytometry17, 294-301 (1994). The remaining erythrocytes in the neutrophilfraction are lysed with sterile water (for 30 seconds) and, afterreinstatement of the isotonicity, the fraction is washed. The cells arefinally inserted (suspended) in HGSS+2% Human Serum Albumin (HSA). Cells(1×10⁴) are incubated with 1 ng/ml ReLPS, 30 ng/ml recombinantLPS-binding protein (r-LPS) and a sample SAP in a total volume of 100 μlfor 30 minutes at 37° C. The samples are subsequently placed in aluminometer (Berthold; Autolumat LB953), whereafter via automaticinjection luminol (180 μM) and 1 μM fMLP(formylmethyonyl-leucyl-phenylalanine) are added as stimulus to eachtube. The luminescence response is measured continuously for 15 minutesand the results expressed as total luminescence response (area under thegraph). See also Weersink et al., Immunology 83, 617-623 (1994).

4.2 Results

1. FIG. 3 shows the inhibition of the LPS “priming” of neutrophils bySAP. In the graph the luminescence counts are plotted against themeasurement time (in minutes). This shows that the activation ofmeutrophils by LPS is greatly reduced by SAP. This is a strongindication that the toxicity of LPS can also be reduced in vivo by SAP.

Example 5 Inhibition of endotoxin-induced sepsis in vivo

5.1 Materials and method. In vivo protection experiments are performedin a peritonitis model in the mouse (Cross et al., Infect. and Immunity61, 2741-2747 (1993)) with ReLPS or intact Gram-negative bacteria. TheSAP preparation is incubated beforehand with ReLPS or O18K5 bacteria(Cross et al., supra) and subsequently administered intraperitoneally toBALB/c mice in a 3xLD₅₀ dose. Survival was followed for 7 days.

8 1 13 PRT Homo sapiens Fragment of the Serum Amyloid P component,peptides 27-39 1 Glu Lys Pro Leu Gln Asn Phe Thr Leu Cys Phe Arg Ala 1 510 2 6 PRT Artificial Sequence Fragment of the Serum Amyloid Pcomponent, peptides 33-38 2 Phe Thr Leu Cys Phe Arg 1 5 3 8 PRTArtificial Sequence Fragment of the Serum Amyloid P component, peptides32-39 3 Asn Phe Thr Leu Cys Phe Arg Ala 1 5 4 8 PRT Artificial SequenceFragment of the Serum Amyloid P component, peptide 30-37 4 Leu Gln AsnPhe Thr Leu Cys Phe 1 5 5 8 PRT Artificial Sequence Fragment of theSerum Amyloid P component, peptide 29-36 5 Pro Leu Gln Asn Phe Thr LeuCys 1 5 6 8 PRT Artificial Sequence Fragment of the Serum Amyloid Pcomponent, peptide 31-38 6 Gln Asn Phe Thr Leu Cys Phe Arg 1 5 7 8 PRTArtificial Sequence Fragment of the Serum Amyloid P component, peptide28-35 7 Lys Pro Leu Gln Asn Phe Thr Leu 1 5 8 8 PRT Artificial SequenceFragment of the Serum Amyloid P component, peptide 27-34 8 Glu Lys ProLeu Gln Asn Phe Thr 1 5

What is claimed is:
 1. A method for treating an animal or patient inneed of lipopolysaccharide neutralization, comprising administering inunit dosage form an amount of a peptide, said peptide comprising SerumAmyloid P component or at least one endotoxin-binding fragment thereof,effective to neutralize lipopolysaccharide in an animal or patient thustreated.
 2. A method for treating an animal or patient in need oftreatment for sepsis, comprising administering in unit dosage form anamount of a peptide, said peptide comprising Serum Amyloid P componentor at least one endotoxin-binding fragment thereof, effective to treatsepsis in an animal or patient on which said method is practiced.
 3. Amethod for treating an animal or patient in need of lipopolysaccharideneutralization, comprising administering in unit dosage form an amountof a peptide, said peptide comprising at least a portion of the sequenceof amino acids 27-39 (SEQ ID NO:1) of Serum Amyloid P component,effective to neutralize lipopolysaccharide in an animal or patient thustreated.
 4. The method according to claim 3 wherein said peptide has asequence selected from the group consisting of:Glu-Lys-Pro-Leu-Gln-Asn-Phe-Thr-Leu-Cys-Phe-Arg-Ala (SEQ ID NO:1),further designated as PEP 27-39; Phe-Thr-Leu-Cys-Phe-Arg (SEQ ID NO:2),further designated as PEP 33-38; Asn-Phe-Thr-Leu-Cys-Phe-Arg-Ala (SEQ IDNO: 3), further designated as PEP 32-39; Leu-Gln-Asn-Phe-Thr-Leu-Cys-Phe(SEQ ID NO:4), further designated as PEP 30-37; andPro-Leu-Gln-Asn-Phe-Thr-Leu-Cys (SEQ ID NO:5), further designated as PEP29-36.
 5. A method for treating an animal or patient in need oftreatment for sepsis, comprising administering in unit dosage form anamount of a peptide, said peptide comprising at least a portion of thesequence of amino acids 27-39 (SEQ ID NO:1) of Serum Amyloid Pcomponent, effective to treat sepsis in an animal or patient on whichsaid method is practiced.
 6. The method according to claim 5 whereinsaid peptide has a sequence selected from the group consisting ofGlu-Lys-Prop-Leu-Gln-Asn-Phe-Thr-Leu-Cys-Phe-Arg-Ala (SEQ ID NO:1),further designated as PEP 27-39; Phe-Thr-Leu-Cys-Phe-Arg (SEQ ID NO:2),further designated as PEP 33-38; Asn-Phe-Thr-Leu-Cys-Phe-Arg-Ala (SEQ IDNO: 3), further designated as PEP 32-39; Leu-Gln-Asn-Phe-Thr-Leu-Cys-Phe(SEQ ID NO:4), further designated as PEP 30-37; andPro-Leu-Gln-Asn-Phe-Thr-Leu-Cys (SEQ ID NO:5), further designated as PEP29-36.